3-Dimensional nano-fibre scaffold for ex vivo expansion of cord blood haematopoietic stem cells

Abstract

Cord blood (CB) haematopoietic stem cell (HSC) is an alternative source of HSC transplantation. The limited cell number greatly restricts their clinic-scale therapeutic applications. The objective of this study was an ex vivo expansion of CB HSCs in a new three-dimensional polycaprolactone nano-scaffold coated with fibronectin (FN). First, we isolated CB CD34+ cells and cultured 10 days in presence of growth factors. The evaluation was performed by qRT-PCR, flow cytometry and clonogenicity. 3D PCL nano-scaffold coated with FN produced significantly higher total nucleated cells and CD34+ cells (p < .05) and also had significantly higher homing and self-renewality genes than 2D cell culture and before expansion (p < .05). The expression of CXCR-4, VLA-4, VLA-5 and LFA-1, and also HOXB-4, HOXA-9, BMI-1 and hTERT genes was higher in 3D than 2D. The CD13, CD14, CD33, CD34 and CD45 markers were significantly higher and CD2, CD3 and CD19 markers were significantly lower in 3D scaffold than 2D cell culture (p < .05). The type and number of colonies in 2D culture were lower than 3D culture medium (p > .05). 3D PCL nano-scaffold coated with FN could better keep specifications homing and self renewality of CB HSCs after expansion.

Keywords:

• Haematopoietic stem cell
• cord blood haematopoietic stem cell expansion
• haematopoietic stem cell transplantation
• cord blood transplantation
• tissue engineering
• 3D culture

Acknowledgements

We would like to thank Dr. Zarrabi, Chief Director of Royan Stem Cell Technology Company (Royan Cord Blood Bank), and Mr. Janzamin for his assistance with flow cytometry.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by the Deputy of Research Affairs and Faculty of Medical Sciences [52/2767] at Tarbiat Modares University.